ROZA-XL, an improved FRET based biosensor with an increased dynamic range for visualizing zeta associated protein 70 kD (ZAP-70) tyrosine kinase activity in live T cells

Biochemical and Biophysical Research Communications
Sophie CadraAnnemarie Coffman Lellouch

Abstract

Genetically encoded FRET based biosensors allow one to visualize the spatial and temporal evolution of specific enzyme activities in live cells. We have previously reported the creation of a FRET based biosensor specific for Zeta-Associated Protein -70 kD (ZAP-70) (Randriamampita et al., 2008), a Syk family protein tyrosine kinase. ZAP-70 is essential for early T cell receptor (TCR) signaling events, T lymphocyte development and has also been implicated in integrin mediated T lymphocyte migration. In order to facilitate the study of ZAP-70 kinase activity during dynamic phenomena such as immunological synapse formation or cell migration, we have designed and prepared a second generation of ZAP-70 specific biosensors. Here we describe a novel biosensor named ROZA-XL, that displays a 3-4 times greater dynamic range than its predecessor and possesses a robust baseline FRET value when expressed in the Jurkat human T cell line. We demonstrate that the robust behavior of this biosensor allows for rapid analysis of TCR mediated of ZAP-70 kinase activity at a single cell level, as shown in a simple end point assay in which ROZA-XL expressing cells are allowed to interact with stimulatory anti-CD3epsilon coated coverslips.

References

Dec 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·A Y TingR Y Tsien
Oct 3, 2002·The Journal of Cell Biology·Stephen C BunnellLawrence E Samelson
Dec 4, 2002·Nature Reviews. Molecular Cell Biology·Jin ZhangRoger Y Tsien
Feb 8, 2005·Nature Biotechnology·Annalee W Nguyen, Patrick S Daugherty
Sep 19, 2008·Proceedings of the National Academy of Sciences of the United States of America·Mingxing OuyangYingxiao Wang
Mar 18, 2009·Immunological Reviews·Byron B Au-YeungArthur Weiss
Aug 12, 2009·Molecular and Cellular Biology·R G BakerG A Koretzky
May 11, 2010·Cold Spring Harbor Perspectives in Biology·Haopeng WangArthur Weiss
Mar 6, 2012·Journal of Cell Science·Alexander A SmoligovetsJay T Groves
Nov 15, 2012·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Rebekah R Bartelt, Jon C D Houtman
Oct 30, 2013·Biotechnology Journal·Clotilde Randriamampita, Annemarie C Lellouch
Aug 8, 2014·Biophysical Journal·Omer DushekVahid Shahrezaei

❮ Previous
Next ❯

Citations

Oct 27, 2015·Sensors·Bernhard HochreiterJohannes A Schmid
Jul 8, 2016·Annals of Biomedical Engineering·Kaitao LiCheng Zhu
Mar 20, 2018·Cell Structure and Function·Gembu MaryuKazuhiro Aoki
Jul 29, 2019·The Journal of Membrane Biology·Laurent Limozin, Pierre-Henri Puech
Oct 20, 2020·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Xinyuan ZhangKeyue Shen
Jun 7, 2021·Current Opinion in Cell Biology·Kendelle J MurphyPaul Timpson

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adhesion Molecules in Health and Disease

Cell adhesion molecules are a subset of cell adhesion proteins located on the cell surface involved in binding with other cells or with the extracellular matrix in the process called cell adhesion. In essence, cell adhesion molecules help cells stick to each other and to their surroundings. Cell adhesion is a crucial component in maintaining tissue structure and function. Discover the latest research on adhesion molecule and their role in health and disease here.

Cell Migration

Cell migration is involved in a variety of physiological and pathological processes such as embryonic development, cancer metastasis, blood vessel formation and remoulding, tissue regeneration, immune surveillance and inflammation. Here is the latest research.

Advanced Imaging of Cellular Signaling

Cell signaling is a vital mechanism for communication within cells and outside with the environment. Several different signaling pathways have been found and advanced imaging techniques are being developed to visualize the molecules involved in these signaling pathways. Find the latest research in advanced imaging of cellular signaling here.